Cell-specific translational profiling in acute kidney injury
Cell-specific translational profiling in acute kidney injury
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DOI：
10.1172/jci72126
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发表时间：
2014-03-01
影响因子：
15.9
通讯作者：
McMahon, Andrew R.
中科院分区：
文献类型：
--
作者：
Liu, Jing;Krautzberger, A. Michaela;McMahon, Andrew R.
急性肾损伤（AM）会导致肾功能突然丧失，从而导致大量发病率和死亡率。相当大的努力已经走向识别的诊断生物标志物和分析AM相关的分子事件，但是，大多数研究都采用了器官范围的方法，并没有阐明AM病理生理学中涉及的不同细胞类型之间的相互作用。为了更好地表征AM相关的分子和细胞事件，我们开发了一种小鼠系，能够识别特定细胞类型中的翻译谱。该策略依赖于EGFP标记的LlOa核糖体蛋白亚基的CRE重组酶依赖性活化，其允许表达CRE的细胞中mRNA群体的翻译核糖体亲和纯化（TRAP）。将该小鼠系与细胞类型特异性CRE驱动系相结合，我们在AM的缺血再灌注损伤（Jill）模型中鉴定了不同的细胞反应。IRI后24小时，在肾单位、肾间质细胞群、血管内皮和巨噬细胞/单核细胞中发现了不同的翻译特征。此外，TRAP捕获了已知的IRI相关标志物，验证了这种方法。生物学功能注释，经典途径分析，并在原位分析确定的反应基因提供了深入了解细胞特异性损伤的签名。我们的研究提供了AM中早期损伤相关分子事件的深入，基于细胞的观点，并记录了一种多功能的遗传工具，用于监测疾病建模中的细胞特异性和时间特异性生物学过程。
Acute kidney injury (AM) promotes an abrupt loss of kidney function that results in substantial morbidity and mortality. Considerable effort has gone toward identification of diagnostic biomarkers and analysis of AM-associated molecular events; however, most studies have adopted organ-wide approaches and have not elucidated the interplay among different cell types involved in AM pathophysiology. To better characterize AM-associated molecular and cellular events, we developed a mouse line that enables the identification of translational profiles in specific cell types. This strategy relies on CRE recombinase-dependent activation of an EGFP-tagged LlOa ribosomal protein subunit, which allows translating ribosome affinity purification (TRAP) of mRNA populations in CRE-expressing cells. Combining this mouse line with cell type-specific CRE-driver lines, we identified distinct cellular responses in an ischemia reperfusion injury (Jill) model of AM. Twenty-four hours following IRI, distinct translational signatures were identified in the nephron, kidney interstitial cell populations, vascular endothelium, and macrophages/monocytes. Furthermore, TRAP captured known IRI-associated markers, validating this approach. Biological function annotation, canonical pathway analysis, and in situ analysis of identified response genes provided insight into cell-specific injury signatures. Our study provides a deep, cell-based view of early injury-associated molecular events in AM and documents a versatile, genetic tool to monitor cell-specific and temporal-specific biological processes in disease modeling.